碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 103 / Progranulin (PGRN), a multi-functional gene, can regulate cell proliferation, survival, wound healing, embryonic development, and inflammation. PGRN consists of multiple Granulin (GRN) units. In this study, we identified PGRN gene in white shrimp (Litopenaeus vannamei), designated Lvpgrn, which is frist discovered in Crustacea. We used RACE-PCR to obtain the full-length cDNA of Lvpgrn from white shrimp haemocytes. The full-length cDNA of Lvpgrn was 2640 bp in length, consisting of a 5'-untranslated region of 137 bp, an ORF of 1719 bp and a 3'-untranslated region of 784 bp. The ORF encodes a protein of 572 amino acid residues, composed of a signal peptide and six GRN units. Expression profiling by real-time PCR (Q-PCR) revealed that Lvpgrn transcripts could be detected in all tissues examined and mainly expressed in intestine, lymphoid organ, heart and nerve. After WSSV infection, Lvpgrn transcripts were significantly increased at 36 hpi and at 48 hpi were increased to 7 times in haemocytes, equal to the expression level of lymphoid organ of uninfected white shrimp. In lymphoid organ, at 48 hpi Lvpgrn transcripts were increased to almost 3 times higher than that in uninfected white shrimp. And the expression profile of LvDscam is similar to Lvpgrn in haemocytes. After WSSV infection, vp28, the envelope protein gene, was significantly increased after 36 hpi in haemocytes; in lymphoid organ, after 24 hpi vp28 transcripts were significantly increased. It can show that the expression levels of Lvpgrn transcripts can be induced in immune-related tissue, haemocytes and lymphoid organ, at late stage of WSSV infection. In the RNA interference experiments, Lvpgrn expression was silenced by injection 1 μg/g shrimp Lvpgrn dsRNA, the shrimp died within 24 h after injection (data not shown). The data showed that the survival rate decreased rapidly from 24 hpi to 72 hpi in high concentration (0.5 μg/g shrimp) and declined in a rush from 36 hpi to 48 hpi in low concentration (0.25 μg/g shrimp) and final survival rates were 25% and 60%, respectively, whereas injection of 0.5 μg/g shrimp GFP dsRNA had 100% survival. The reports indicated that Lvpgrn play an essential role in white shrimp survival. The results of Q-PCR displayed that the mRNA level of Lvpgrn was knocked down by approximately 21% and 75% in haemocytes and by approximately 72% and 73% in lymphoid organ at 1 dpi and 3 dpi, compare with injection of GFP dsRNA at 1 dpi. Although we recognize that Lvpgrn will be regulated by WSSV infection and plays an essential role in shrimp survival, the function and relative pathway of Lvpgrn has not yet to be demonstrated and has not been reported up to now.
| Identifer | oai:union.ndltd.org:TW/103NTOU5086037 |
| Date | January 2015 |
| Creators | Lu, Chia-Wen, 盧佳彣 |
| Contributors | Gong, Hong-Yi, 龔紘毅 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 63 |
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